Journal: Acta neuropathologica communications

Article Title: Trichuris suis induces human non-classical patrolling monocytes via the mannose receptor and PKC: implications for multiple sclerosis.

doi: 10.1186/s40478-015-0223-1

Figure Lengend Snippet: Fig. 6 PKC signals downstream upon TsSP-MR interaction. Quantification of flow cytometry analysis of the expression levels of (a) CCR2, (b) LFA-1 avidity and (c) LFA-1 affinity on human monocytes. These cells were cultured in the presence or absence of TsSP (40 μg/ml, 16 h), the PKC inhibitor Bisindolylmaleimide I (GF109203, 2 μM) and blocking antibodies for the MR or its isotype control (both 10 μg/ml). (b) and (c) are expressed as the percentage of avidity/affinity compared to total LFA-1. Cell supernatants were used to determine the levels of (d) TNF-α and (e) IL-10 secretion by enzyme-linked immunosorbent assay analysis. f PKC activation was determined by Western Blotting using phosphoPKC antibodies compared to GAPDH expression and images were subsequently quantified using ImageJ. Experiments were performed in triplicate using cells derived from 4 different human donors and the results are presented as the mean (a-c); the mean percentage (d, e) or the mean PKC phosphorylation +/−SEM. *p < 0.05, **p < 0.01, ***p < 0.001 as determined by ANOVA and Students t test

Article Snippet: The following primary antibodies (in TSM/0,05 % Tween/5 % BSA) were used: PhosphoPKC (Cell signaling, Beverly, MA, USA, #9371S) and GAPDH (Santa Cruz Biotechnology, Heidelberg, Germany, #sc-32233) as a loading control.

Techniques: Flow Cytometry, Expressing, Cell Culture, Blocking Assay, Control, Enzyme-linked Immunosorbent Assay, Activation Assay, Western Blot, Derivative Assay, Phospho-proteomics

Journal: Andrology

Article Title: Catch-up fat in male adults induces low testosterone and consequently promotes metabolic abnormalities and cognitive impairment.

doi: 10.1111/andr.13177

Figure Lengend Snippet: FIGURE 3 Alteration of cognition function, insulin signal pathway, and NR2A/2B expression in the cerebral cortex of male catch-up fat in adult (CUFA) rats. Comparison of the numbers of trials required to reach the criterion of six consecutive correct in the different stages of ASST (A), the relative mRNA expression level of IRS-1 (B), insulin-stimulated AKT phosphorylation at Ser473 (C), the relative protein expression level of AR (D), relative mRNA expression (E) and protein expression (F) level of NR2A, relative mRNA expression (G) and protein expression (H) level of NR2B between NC and RN groups at 12 w. *P < 0.05, **P < 0.01, ***P < 0.005. Abbreviations: NC, normal chow controls; RN, caloric restriction and refeeding with normal chow (CUFA rats); ASST, Attentional set-shifting task; SD, Simple discrimination; CD, Compound discrimination; R1, First reversal; IDS, Intra-dimensional set-shifting; R2, Second reversal; EDS, Extradimensional set-shifting; R3, Third reversal; IRS-1, Insulin receptor substrate 1, AKT, serine/threonine kinase 1; AR, androgen receptor; NR2A, N-methyl-d-aspartate receptors subtype 2A, NR2B, N-methyl-d-aspartate receptors subtype 2B

Article Snippet: Proteins extracts from the cerebral cortex of rats were prepared and immunoprecipitation was performed as reported previously.3 The antibodies used for immunoblot assays were anti-PKB (Protein kinase B)/Akt (1: 1000, Cell Signaling Technology), anti-phosphoPKB/Akt (Ser473) (1: 1000, Cell Signaling Technology), anti-NR2A (1:2000, Proteintech Group, Inc.), anti-NR2B (1:2000, Proteintech Group, Inc.), anti-AR (androgen receptor) (Proteintech Group, Inc), and anti-GAPDH (1:3000, Gene Tex).

Techniques: Expressing, Comparison, Phospho-proteomics

Journal: Andrology

Article Title: Catch-up fat in male adults induces low testosterone and consequently promotes metabolic abnormalities and cognitive impairment.

doi: 10.1111/andr.13177

Figure Lengend Snippet: FIGURE 6 Testosterone supplementation attenuated cognitive impairment as well as the impaired insulin signal pathway and suppressed NA2A/2B expression in male catch-up fat in adults (CUFA) rats. Comparison of the number of trials required to achieve the criterion of six consecutive correct in the different stages of ASST (A), the relative mRNA expression level of IRS-1 (B), insulin-stimulated AKT phosphorylation at Ser473 in brain tissue (C), the relative protein expression level of AR (D), relative mRNA expression (E) and protein expression (F) level of NR2A, relative mRNA expression (G) and protein expression (H) level of NR2B between RN and RT groups at 12w. *p < 0.05, **p < 0.01, ***p < 0.005. Abbreviations: RN, caloric restriction and refeeding with normal chow (CUFA rats); RT: CUFA rat supplemented with testosterone; ASST, Attentional set-shifting task; SD, Simple discrimination; CD, Compound discrimination; R1, First reversal; IDS, Intra-dimensional set-shifting; R2, Second reversal; EDS, Extradimensional set-shifting; R3, Third reversal; IRS-1, insulin receptor substrate 1, AKT, serine/threonine kinase 1; AR, androgen receptor; NR2A, N-methyl-d-aspartate receptors subtype 2A, NR2B, N-methyl-d-aspartate receptors subtype 2B

Article Snippet: Proteins extracts from the cerebral cortex of rats were prepared and immunoprecipitation was performed as reported previously.3 The antibodies used for immunoblot assays were anti-PKB (Protein kinase B)/Akt (1: 1000, Cell Signaling Technology), anti-phosphoPKB/Akt (Ser473) (1: 1000, Cell Signaling Technology), anti-NR2A (1:2000, Proteintech Group, Inc.), anti-NR2B (1:2000, Proteintech Group, Inc.), anti-AR (androgen receptor) (Proteintech Group, Inc), and anti-GAPDH (1:3000, Gene Tex).

Techniques: Expressing, Comparison, Phospho-proteomics